Aliphatic Quaternary Ammonium Functionalized Nanogels for Gene Delivery.

Huaiying Zhang,Damla Keskin,Patrick Van Rijn,Inge S Zuhorn,Willy H De Haan-Visser,Guangyue Zu

doi:10.3390/pharmaceutics13111964

Huaiying Zhang, Damla Keskin + Show 4 more

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https://doi.org/10.3390/pharmaceutics13111964

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Abstract

Gene therapy is a promising treatment for hereditary diseases, as well as acquired genetic diseases, including cancer. Facing the complicated physiological and pathological environment in vivo, developing efficient non-viral gene vectors is needed for their clinical application. Here, poly(N-isopropylacrylamide) (p(NIPAM)) nanogels are presented with either protonatable tertiary amine groups or permanently charged quaternized ammonium groups to achieve DNA complexation ability. In addition, a quaternary ammonium-functionalized nanogel was further provided with an aliphatic moiety using 1-bromododecane to add a membrane-interacting structure to ultimately facilitate intracellular release of the genetic material. The ability of the tertiary amine-, quaternized ammonium-, and aliphatic quaternized ammonium-functionalized p(NIPAM) nanogels (i.e., NGs, NGs-MI, and NGs-BDD, respectively) to mediate gene transfection was evaluated by fluorescence microscopy and flow cytometry. It is observed that NGs-BDD/pDNA complexes exhibit efficient gene loading, gene protection ability, and intracellular uptake similar to that of NGs-MI/pDNA complexes. However, only the NGs-BDD/pDNA complexes show a notable gene transfer efficiency, which can be ascribed to their ability to mediate DNA escape from endosomes. We conclude that NGs-BDD displays a cationic lipid-like behavior that facilitates endosomal escape by perturbing the endosomal/lysosomal membrane. These findings demonstrate that the presence of aliphatic chains within the nanogel is instrumental in accomplishing gene delivery, which provides a rationale for the further development of nanogel-based gene delivery systems.

Highlights

Gene therapy represents a promising approach for the treatment of a wide variety of human diseases, such as Duchenne muscular dystrophy, Parkinson’s disease, Huntington’s disease, and cancers [1,2,3,4]
The tertiary amines were alkylated by methyl iodide (MI) or 1-bromododecane (BDD), creating the quaternized nanogels
Investigation by atomic force microscopy showed that all three nanogels, i.e., NGs, NGs-MI, and Nanogel with 1-Bromododecane (NGs-BDD), possessed a spherical shape and a homogeneous particle size (Figure 2A)

Summary

Introduction

Gene therapy represents a promising approach for the treatment of a wide variety of human diseases, such as Duchenne muscular dystrophy, Parkinson’s disease, Huntington’s disease, and cancers [1,2,3,4]. The most extensively used delivery vectors are viral vectors, which enable the efficient delivery of genes. Concerns with the use of viruses, including the high production cost, and risk of immunogenicity and induced oncogenic transformations have led researchers to search for non-viral gene vectors [5,6]. While cationic lipids bearing a quaternary ammonium group are successfully used for in vitro gene delivery, quaternary ammonium cationic polymers generally show reduced transfection efficiency and increased cytotoxicity [15,16].

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